Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Sakai, H.
    Mutagenesis of the active site lysine 221 of the pyruvate kinase from Bacillus stearothermophilus (2005), J. Biochem., 137, 141-145.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.7.1.40 D-ribose 5-phosphate 0.1 mM, allosteric activator Geobacillus stearothermophilus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.40 expressed in Escherichia coli XL-1 Blue cells Geobacillus stearothermophilus

Protein Variants

EC Number Protein Variants Comment Organism
2.7.1.40 C9S/C268S 75% activity of the wild-type enzyme Geobacillus stearothermophilus
2.7.1.40 C9S/C268S/K221C 10000 times less active than the wild type enzyme Geobacillus stearothermophilus
2.7.1.40 K221C 10000 to 100000 times less active than the wild-type enzyme Geobacillus stearothermophilus
2.7.1.40 K221D 10000 to 100000 times less active than the wild-type enzyme Geobacillus stearothermophilus
2.7.1.40 K221L 10000 to 100000 times less active than the wild-type enzyme Geobacillus stearothermophilus
2.7.1.40 K221R 10000 to 100000 times less active than the wild-type enzyme Geobacillus stearothermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.40 0.113
-
 phosphoenolpyruvate mutant enzyme C9S/C268S, in the presence of 0.1 mM ribose 5'-phosphate, in 50 mM imidazole-HCl buffer, pH 7.2, 50 mM KCl, 7 mM MgCl2, 0.12 mM NADH, 0.02 mg/ml lactate dehydrogenase, 2 mM PEP, and 4 mM ADP, at 30°C Geobacillus stearothermophilus
2.7.1.40 0.186
-
 phosphoenolpyruvate wild type enzyme, in the presence of 0.1 mM ribose 5'-phosphate, in 50 mM imidazole-HCl buffer, pH 7.2, 50 mM KCl, 7 mM MgCl2, 0.12 mM NADH, 0.02 mg/ml lactate dehydrogenase, 2 mM PEP, and 4 mM ADP, at 30°C Geobacillus stearothermophilus

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.40 Geobacillus stearothermophilus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.40 Butyl-Toyopearl 650S column chromatography and MonoQ HR10/10 column chromatography Geobacillus stearothermophilus

Renatured (Commentary)

EC Number Renatured (Comment) Organism
2.7.1.40 dialysis against 0.2 M Tris-HCl, pH 8.6, containing 1 mM EDTA and 0.05 mM dithiothreitol, after denaturation with 4 M guanidine hydrochloride Geobacillus stearothermophilus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.40 additional information
-
 555 units/mg, wild type enzyme, at 30°C Geobacillus stearothermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.40 ADP + phosphoenolpyruvate
-
 Geobacillus stearothermophilus ATP + pyruvate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.1.40 phosphoenolpyruvate kinase
-
 Geobacillus stearothermophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.40 1736
-
 phosphoenolpyruvate mutant enzyme C9S/C268S, in the presence of 0.1 mM ribose 5'-phosphate, in 50 mM imidazole-HCl buffer, pH 7.2, 50 mM KCl, 7 mM MgCl2, 0.12 mM NADH, 0.02 mg/ml lactate dehydrogenase, 2 mM PEP, and 4 mM ADP, at 30°C Geobacillus stearothermophilus
2.7.1.40 3204
-
 phosphoenolpyruvate wild type enzyme, in the presence of 0.1 mM ribose 5'-phosphate, in 50 mM imidazole-HCl buffer, pH 7.2, 50 mM KCl, 7 mM MgCl2, 0.12 mM NADH, 0.02 mg/ml lactate dehydrogenase, 2 mM PEP, and 4 mM ADP, at 30°C Geobacillus stearothermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.40 ADP
-
 Geobacillus stearothermophilus